A flat embedding method for transmission electron microscopy reveals an unknown mechanism of tetracycline
Artikel i vetenskaplig tidskrift, 2021

Transmission electron microscopy of cell sample sections is a popular technique in microbiology. Currently, ultrathin sectioning is done on resin-embedded cell pellets, which consumes milli- to deciliters of culture and results in sections of randomly orientated cells. This is problematic for rod-shaped bacteria and often precludes large-scale quantification of morphological phenotypes due to the lack of sufficient numbers of longitudinally cut cells. Here we report a flat embedding method that enables observation of thousands of longitudinally cut cells per single section and only requires microliter culture volumes. We successfully applied this technique to Bacillus subtilis, Escherichia coli, Mycobacterium bovis, and Acholeplasma laidlawii. To assess the potential of the technique to quantify morphological phenotypes, we monitored antibiotic-induced changes in B. subtilis cells. Surprisingly, we found that the ribosome inhibitor tetracycline causes membrane deformations. Further investigations showed that tetracycline disturbs membrane organization and localization of the peripheral membrane proteins MinD, MinC, and MreB. These observations are not the result of ribosome inhibition but constitute a secondary antibacterial activity of tetracycline that so far has defied discovery.

Peptidoglycans

Cell shape

Plasma protein Z

Författare

Michaela Wenzel

Universiteit Van Amsterdam

Chalmers, Biologi och bioteknik, Kemisk biologi

Swammerdam Institute for Life Sciences

Marien P. Dekker

Vrije Universiteit Amsterdam

Biwen Wang

Swammerdam Institute for Life Sciences

Maroeska J. Burggraaf

Universiteit Van Amsterdam

Wilbert Bitter

Universiteit Van Amsterdam

Vrije Universiteit Amsterdam

Jan R.T. van Weering

Vrije Universiteit Amsterdam

Leendert W. Hamoen

Swammerdam Institute for Life Sciences

Communications Biology

23993642 (eISSN)

Vol. 4 1 306-

Ämneskategorier

Cellbiologi

Cell- och molekylärbiologi

Medicinsk bioteknologi (med inriktning mot cellbiologi (inklusive stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci)

DOI

10.1038/s42003-021-01809-8

PubMed

33686188

Mer information

Senast uppdaterat

2021-04-22